Pharmaceutical composition

ABSTRACT

The present invention relates to pharmaceutical compositions having improved stability.

The present invention relates to pharmaceutical compositions, forexample pharmaceutical compositions for the treatment of post-partumhaemorrhage (PPH) or other medical applications. In particular, itrelates to pharmaceutical compositions having improved stability, forexample at room or ambient temperature.

Postpartum haemorrhage (PPH) is one of the leading causes ofpregnancy-related mortality and severe morbidity in developing countriesas well as in the industrialized world. It is a potentiallylife-threatening condition which is demonstrated by about 140,000 deathsannually, one every four minutes, the vast majority among women who donot have access to adequate obstetric healthcare. Although the problemis numerically important, not all regions of the industrialized worldare similarly affected and in Europe, the rates of death associated withmaternal haemorrhage vary widely from one country to another, Apopulation based survey in 11 regions within European countries showedthat rates of severe haemorrhage ranged from 0.1% to 0.9% of pregnancies(MOMS-B group, 1999). There are good reasons to believe that differencesin clinical practice may be of great importance to the differences inmorbidity/mortality. In the United Kingdom, the Confidential Enquiryinto Maternal Deaths, covering the years 1994-1996, showed that afteraction had been taken in all maternity units to establish guidelines formanagement of third stage and postpartum haemorrhage, there were nodeaths from haemorrhage in uncomplicated vaginal births (Department ofHealth et al, 1998). This achievement supports the assumption thatclinical management has a key role in prevention of severe maternalhaemorrhage.

PPH is difficult to manage because evaluation of blood loss in thedelivery unit is unreliable. Action is often taken in response to thedevelopment of maternal signs such as hypotension or malaise rather thanon the basis of estimated blood loss. The delayed action is responsiblefor many cases of severe haemorrhage, and immediate surgery may berequired because time spent using other treatment methods would bedangerous for the patient. These considerations speak in favour of theapplication of a policy of routine prophylactic administration of auterotonic to all parturients. The case for such a policy is augmentedby the fact that uterine atony is the most significant contributor toPPH. Uterine atony is a loss of tone in the uterine musculature.Normally, contraction of the uterine muscle compresses the vessels andreduces flow. This increases the likelihood of coagulation and preventsbleeds. Thus, lack of uterine muscle contraction can cause an acutehaemorrhage. Clinically, 75-80% of postpartum hemorrhages are due touterine atony.

Reviews have shown compelling evidence in support of routineprophylactic administration of a uterotonic, either in isolation or aspart of the entity Active Management of the Third Stage of Labour(AMTSL); AMTSL is usually defined as an intervention with threecomponents: a prophylactic administration of a uterotonic agent, earlycord clamping and controlled cord traction. Further, AMTSL is shown tobe equally effective in “low-risk” and “high-risk” women. Today, the useof a uterotonic drug as prophylaxis in all vaginal hospital deliveriesto prevent severe haemorrhage is routine clinical management in most ofEurope, and the practice is increasing globally.

The uterotonic drugs currently available are oxytocin, ergometrine,Syntometrine® (a combination of oxytocin and ergometrine) andmisoprostol. However, these are not without disadvantages. Misoprostolis administered orally or vaginally and is less efficacious thaninjectable uterotonics; it is generally recommended that it is not usedwhere injectable uterotonics are available. Syntometrine® is licensed inonly a few countries in Europe. It is possibly more effective thanoxytocin alone but is associated with more side effects, particularlynausea and vomiting. Further, it is unsuitable for use in women withhypertension, preeclampsia and heart disease, thereby reducing itssuitability for routine prophylactic usage. Oxytocin itself has thedisadvantage of a short half-life. Although this can be circumvented byadministration as a continuous intravenous infusion to provide sustaineduterotonic activity, this is more inconvenient than a single injection.If administered as a single bolus dose, whether as intravenous orintramuscular injection, close monitoring of uterine tone is requiredand additional uterotonic medication may be required due to the shorthalf-life.

Carbetocin [(1-desamino-1-monocarba-2(O-methyl)-tyrosine)oxytocin] is along-acting synthetic analogue of oxytocin, with agonist action.Carbetocin (PABAL®, DURATOCIN®) is currently approved for the preventionof uterine atony following delivery of the infant by Caesarean sectionunder epidural or spinal anaesthesia. In its present marketed usage,intravenous administration of carbetocin provides a half-life ofapproximately 40 minutes, which is 4 to 10 times longer than thereported half-life of oxytocin (4 to 10 minutes). However, usingintramuscular injection, carbetocin reaches peak plasma concentrationsin less than thirty minutes and has a bioavailability of 80% (W Rath,European Journal of Obstetrics and Gynecology and Reproductive Biology147 (2009) 15-20). Thus, carbetocin has the potential to be a near idealdrug for routine postpartum haemorrhage prophylaxis, offered in allhospital vaginal deliveries, because it is suitable for bothintramuscular injection and intravenous administration, offeringconvenience and simple implementation; it has quick onset of action; islong-acting, especially compared to oxytocin; is rarely associated withadverse drug reactions; and has excellent tolerability. In publishedclinical trials, carbetocin has shown efficacy similar to oxytocin andSyntometrine® or even a trend towards better efficacy, demonstrated onseveral outcomes: blood loss (measured or estimated), incidence of bloodloss >500 ml, additional use of uterotonic medication or totaluterotonic interventions. Thus, carbetocin should offer an improvementover currently available options for prevention of uterine atony andexcessive bleeding following vaginal delivery. In comparison withoxytocin, its advantage is primarily that it would replace the need forcontinuous infusion or additional uterotonic intervention. In practice,carbetocin has the potential to replace 2-4 hours of postpartum routineinfusion therapy, and/or to improve outcome versus oxytocin bolusinjection. Further, any reduction in additional interventions representsa favourable pharmacoeconomic case for use of carcetocin. In comparisonwith Syntometrine®, its advantage is primarily better tolerability andsafety and lack of important contraindications. In either case,carbetocin is better suited for, and is likely to facilitate theimplementation of, routine prophylactic usage.

The current carbetocin formulation (PABAL® 100 micrograms/mL solutionfor injection, Ferring Pharmaceuticals Limited) is not room temperature(RT) stable and requires refrigerated storage at a temperature of 2-8°C. There is, therefore, a need for a formulation of carbetocin which isroom temperature stable (for example at 25° C. and 60% relativehumidity) for up to two years, allowing, for example, use in ambulances.This would provide an advantage in climatic zone I/II. More importantly,there is a need for a formulation which may be stored unrefrigerated inclimatic zone III/IV (high temperature, e.g. tropical) regions—that ismeeting the temperature and humidity stability requirements for thesezones, for example, a documented long-term temperature stability at 30°C. and relative humidity up to 75%. The climatic zone terminology isused by the FDA and EMEA and is familiar to those skilled in the art.Thus, climatic zone I is a temperate climate; climatic zone II is asubtropical and mediterranean climate; climatic zone III is hot and dry;and climatic zone IV is hot and humid.

According to the present invention there is provided a liquidcomposition (e.g. a liquid pharmaceutical composition) comprisingcarbetocin or a pharmaceutically active salt thereof; wherein the pH ofthe composition is from 5.0 to 6.0. The pH of the composition may befrom 5.1 to 6.0, for example from 5.2 to 6, for example from 5.26 to 6.The pH of the composition may be from 5.15 to 5.75, for example from 5.2to 5.65. The pH of the composition may be from 5.26 to 5.8, for examplefrom 5.26 to 5.75, for example from 5.26 to 5.7, for example from 5.26to 5.65, for example 5.4 to 5.65. Preferably, the liquid composition(e.g. a liquid pharmaceutical composition) comprises carbetocin.

Preferably the composition is an aqueous composition (e.g. an aqueouspharmaceutical composition) comprising carbetocin or a pharmaceuticallyactive salt thereof; wherein the pH of the composition is from 5.0 to6.0. The pH of the composition may be from 5.1 to 6.0, for example from5.2 to 6, for example from 5.26 to 6. The pH of the composition may befrom 5.15 to 5.75, for example from 5.2 to 5.65. The pH of thecomposition may be from 5.26 to 5.8, for example from 5.26 to 5.75, forexample from 5.26 to 5.7, for example from 5.26 to 5.65, for example 5.4to 5.65. It will be appreciated that the composition of the invention ispreferably an aqueous solution. Although water (e.g. water for injectionor WFI) is the preferred solvent, other solvents (mixtures of water withother pharmaceutically acceptable solvents, pharmaceutically acceptablealcohols, etc.) may be used.

The Applicants have found (Examples 1 to 3) that a composition, e.g. apharmaceutical composition, comprising carbetocin or pharmaceuticallyactive salt thereof, and having pH within a defined specific pH range,may be stored at room temperature (e.g. at 25° C. and 60% relativehumidity) for a sustained period (e.g. up to 2 years). The compositionmay also have a long-term temperature stability at 30° C. and 40° C. andrelative humidity up to 75%, and therefore be suitable for use in zoneIII/IV regions without requirement for refrigeration.

The composition may include (comprise) a buffering agent, for example apharmaceutically acceptable buffering agent. Herein, the term bufferingagent is an agent which is capable of driving an acidic or basicsolution to a certain pH state, and then preventing a change from thatstate; in other words a buffering agent is an agent which is added to analready acidic or basic solution to modify the pH and then maintain thepH at the modified level. Generally a buffering agent is a weak acid ora weak base that would be comprised in a buffer solution, and beresponsible for the buffering action seen in these solutions. Thebuffering agent may be, for example, acetic acid, adipic acid, citricacid, maleic acid, succinic acid or phosphate (e.g. sodium phosphate,e.g. sodium phosphate dibasic dihydrate). Preferably, the bufferingagent is succinic acid. The composition may include a single bufferingagent (i.e. not include two or more buffering agents). The compositionmay include two or more buffering agents (e.g. citric acid and (e.g.sodium) phosphate.

In another aspect, the composition may include (comprise) a buffersolution. Herein, the term buffer or buffer solution means a solutionincluding a mixture of a weak acid and its conjugate base or a weak basean its conjugate acid, which has the property that the pH of thesolution changes very little when a small amount of strong acid or baseis added, such that the pH of the buffer (solution) is maintained. Thebuffer (solution) may be, for example, a citrate buffer (solution),comprising citric acid and a citrate (e.g. sodium citrate); a succinatebuffer (solution) comprising succinic acid and a succinate (e.g. sodiumsuccinate), an acetate buffer (solution) comprising acetic acid and anacetate (e.g. sodium acetate); a citrate/phosphate buffer (solution)comprising citric acid and phosphate; or a phosphate buffer (solution)comprising e.g. (monosodium) phosphate and its conjugate base, (disodiumphosphate). A preferred buffer is a succinate buffer. The compositionmay include a single buffer (i.e. not include two or more buffers). Thecomposition may include two or more buffers.

The Applicants have found that inclusion of succinic acid bufferingagent (or use of a succinate buffer) may provide effective roomtemperature stability (e.g. at 25° C. and 60% relative humidity) whilepossibly conferring additional advantages—for example, the use of asuccinic acid buffering agent or succinate buffer may contribute toreduced injection site reactions and associated pain compared with otherbuffered formulations.

The concentration of carbetocin in the liquid (composition e.g. aqueouscomposition) may be from 0.01 to 55 mg/mL, for example 0.01 to 50 mg/mL,for example 0.01 to 10 mg/mL, for example 0.01 to 1.5 mg/mL, preferably0.05 to 0.5 mg/mL, for example 0.1 mg/mL. The concentration ofcarbetocin in the liquid (composition e.g. aqueous composition) may be,for example, 1 mg/mL, 10 mg/mL, 50 mg/mL etc.

The compositions of the invention may further comprise an anti-oxidant.The anti-oxidant may be any anti-oxidant commonly used in the art, forexample any anti-oxidant approved for use as a pharmaceutical excipient.For example, the anti-oxidant may be methionine, EDTA, butylated hydroxytoluene, sodium metabisulfite etc. Preferably the anti-oxidant ispresent in an amount of 0.01% to 10% (w/v), for example 0.05% to 5%(w/v), most preferably 0.08% to 1% (w/v). Preferably the anti-oxidant ismethionine, EDTA, or a combination of methionine and EDTA. For example,the antioxidant may be methionine and present in an amount of 0.1% w/v(or 1 mg/mL— see Example 2).

The composition may further comprise an isotonicity agent. Isotonicityagents, for example mannitol or NaCl, are well known in the art.Preferably the isotonicity agent is present in an amount sufficient toprovide an isotonic composition (solution), for example in an amount of0.01% to 10% (w/v). Preferably the isotonicity agent is mannitol. If theisotonicity agent is mannitol it may be present in an amount of 0.5% to7.5% (w/v), more preferably 4.0% to 5.5% (w/v), for example 5.0% (w/v).If the isotonicity agent is mannitol it may be present in an amount of0.05% to 7.5% (w/v). If the isotonicity agent is NaCl, it may be presentin an amount of 0.05% to 1.2% (w/v), more preferably 0.08% to 1% (w/v),for example 0.9% (w/v). The isotonicity agent may be present in anamount of 0.1 to 100 mg/mL, for example 0.5 to 7 mg/mL, for example 1 to5 mg/mL. For example, if the isotonicity agent is mannitol it may bepresent in an amount of 5 to 75 mg/mL, for example 40 to 55 mg/mL (seee.g. Table 3a). If the isotonicity agent is NaCl it may be present in anamount of 0.5 to 12 mg/mL, for example 8 to 10 mg/mL (see e.g. Table3b), for example 7.5 mg/mL (see Example 6).

The composition may be for any route of drug administration, e.g. oral,rectal, buccal, nasal, vaginal, transdermal (e.g. patch technology);parenteral, intravenous, intramuscular or subcutaneous injection;intracisternal, intravaginal, intraperitoneal, local (powders, ointmentsor drops) or as a buccal or nasal spray. Preferably the composition isan injectable composition or injectable formulation. Injectableformulations can be supplied in any suitable container, e.g. ampoule,vial, pre-filled syringe, injection device (e.g. single use injectiondevice such as that sold under the mark Uniject by Becton Dickinson),injection cartridge, ampoule, (multi-) dose pen and the like. Preferablythe composition is for intramuscular administration (e.g. intramuscularinjection) or intravenous administration (e.g. IV injection).

The composition may include an enhancer, an excipient which enhances theeffective dose (e.g. enhances the effective dose following nasaladministration). The enhancer may be any enhancer commonly used in theart, for example any enhancer approved for use as a pharmaceuticalexcipient. The enhancer may be, for example, methyl-β-cyclodextrin,Polysorbate 80, carboxymethylcellulose or hydroxypropylmethylcellulose.

The compositions of the invention may be for use in (or in themanufacture of medicaments for) the treatment or prevention of uterineatony. The compositions may be for use in the treatment or prevention ofuterine atony following vaginal delivery of the infant. The compositionsmay be for use in the treatment or prevention of uterine atony followingdelivery of the infant by Caesarean section, for example delivery of theinfant by Caesarean section under epidural or spinal anaesthesia. Thecompositions may be for use in the treatment or prevention of uterineatony, for example in a patient who is at risk of developing PPH. Thecompositions may be for use in (or in the manufacture of medicamentsfor) the treatment or prevention of bleeding (e.g. excessive bleeding)following vaginal delivery (of the infant). The compositions of theinvention may be for use as a uteronic formulation. The compositions ofthe invention may be for (e.g. routine) administration following vaginaldelivery of the infant.

According to the present invention in a further aspect there is provideda method of treatment or prevention of uterine atony (for examplefollowing vaginal delivery of the infant or delivery of the infant byCaesarean section, or in a patient who is at risk of developing PPH) ora method of treatment or prevention of excessive bleeding followingvaginal delivery comprising, a step of administration to a patient inneed thereof a composition as set out above.

It is preferred that the compositions of the invention do not include aquaternary amine compound, such as benzalkonium chloride. It ispreferred that the compositions of the invention do not include aparahydroxybenzoate preservative, or a combination ofparahydroxybenzoate preservative with a cosolvent. It is preferred thatthe compositions of the invention have a content of divalent metal ionsof less than 2 mM, for example 0.195 mM or less, for example 0.1 nM orless. It is preferred that compositions of the invention do not includea solubilizer. It is preferred that compositions of the invention do notinclude methyl-β-cyclodextrin.

In another aspect of the invention, there are provided stabilisedformulations of carbetocin or other pharmaceutically active compounds(e.g. other pharmaceutically active peptides or pharmaceutically activesmall molecules). Thus, according to the invention in a further aspectthere is provided a liquid (e.g. aqueous) composition comprising: apharmaceutically active compound or salt thereof; and an anti-oxidant;wherein the pH of the composition is from 5.0 to 6.0. The pH of thecomposition may be from 5.1 to 6.0, for example from 5.2 to 6, forexample from 5.26 to 6. The pH of the composition may be from 5.15 to5.75, for example from 5.2 to 5.65. The pH of the composition may befrom 5.0 to 5.9, for example from 5.1 to 5.9, for example 5.2 to 5.8.The pH of the composition may be from 5.26 to 5.8, for example from 5.26to 5.75, for example from 5.26 to 5.7, for example from 5.26 to 5.65,for example 5.4 to 5.65. The pharmaceutically active compound may becarbetocin. The composition may be for nasal administration. Thepharmaceutically active compound may be a compound having the formula(I) or solvate or pharmaceutically acceptable salts thereof:

wherein: n is selected from 0, 1 and 2; p is selected from 0, 1, 2, 3,4, 5 and 6; R₁ is selected from aryl optionally substituted with atleast one OH, F, Cl, Br, alkyl or O-alkyl substituent; R₂ is selectedfrom R₄, H, alkyl, cycloalkyl, aryl and 5- and 6-membered heteroaromaticring systems; R₃ is selected from H and a covalent bond to R₂, when R₂is R₄, to form a ring structure; R₄ is C₁₋₆ alkylene moiety substitutedwith at least one O-alkyl, S-alkyl or OH substituent; W and X are eachindependently selected from CH₂ and S, but may not both be CH₂; alkyl isselected from C₁₋₆ straight and C₄₋₈ branched chain alkyl and optionallyhas at least one hydroxyl substituent; aryl is selected from phenyl andmono- or poly-substituted phenyl; with the proviso that when R₂ is H, pis 1, R₃ is H, n is 1 and W and X are both S, R₁ is not 4-hydroxyphenyl.In the above and herein, aryl denotes an aromatic group selected fromphenyl and mono- or polysubstituted phenyl; the substituent moieties, ifpresent, may be selected from fluorine (F), chlorine (CI) and bromine(Br) atoms and alkyl, hydroxy (—OH), alkoxy (—O-alkyl) and alkylthio(—S-alkyl). Preferably the pharmaceutically active compound is acompound according to formula (I) above with the proviso that when R₂ isH, p is 0, R₃ is H, n is 1 and W and X are both S, R₁ is not4-hydroxyphenyl. These compounds, their medical uses and their methodsof their preparation are disclosed in WO2009/122285 (InternationalPatent Application No. PCT/IB2009/005351) of Ferring B.V.

Preferably the pharmaceutically active compound iscarba-1-[4-FBzIGly⁷]dOT, wherein 4-FBzIGly is N-(4-fluorobenzyl)glycine.Carba-1-[4-FBzIGly⁷]dOT is an oxytocin analogue also known as FE 202767.FE 202767 has the structure of formula (II):

FE 202767 is a selective oxytocin receptor agonist being evaluated forclinical use. FE 202767 of formula (II), some medical uses thereof, andmethods of its preparation are disclosed in WO2009/122285. FE202767 isExample 49 of WO2009/122285 (International Patent Application No.PCT/IB2009/005351) of Ferring B.V.

Peptidic oxytocin agonists such as those disclosed in WO2009/122285 areexpected to be delivered by the intranasal route (i.e. intranasaladministration). Intranasal formulations of this type are generallycontained in (and administered using) spray devices, where the drugremains in solution (e.g. at a concentration of active compound of0.05-2 mg/mL) under inert atmosphere for an extended period of time (upto 2 years). There is therefore a need for room temperature stableformulations of oxytocin agonists/analogues such ascarba-1-[4-FBzIGly⁷]dOT (FE 202767), e.g. to avoid requirement forrefrigeration.

The composition may comprise a buffering agent, for example acetic acid,adipic acid, citric acid, maleic acid, succinic acid or (e.g. sodium)phosphate. The composition may include a single buffering agent. Thecomposition may include more than one buffering agent (e.g. may comprisecitric acid and (e.g. sodium) phosphate). The composition may comprise abuffer (solution), for example, a citrate buffer (solution), comprisingcitric acid and a citrate (e.g. sodium citrate); a succinate buffer(solution) comprising succinic acid and a succinate (e.g. sodiumsuccinate), an acetate buffer (solution) comprising acetic acid and anacetate (e.g. sodium acetate); a citrate/phosphate buffer (solution)comprising citric acid and phosphate; or a phosphate buffer (solution).It is preferred, however, that if the pharmaceutically active compoundis 1-deamino-8-D-arginine vasopressin (desmopressin) the buffering agentis not malic acid (and/or the composition does not include malic acidbuffer).

The concentration of the pharmaceutically acceptable compound in theliquid (aqueous) composition may be, for example, 0.01 to 5 mg/mL.

The concentration of carbetocin in the liquid (aqueous) composition maybe from 0.01 to 55 mg/mL, for example 0.01 to 50 mg/mL, for example 0.01to 10 mg/mL, for example 0.01 to 1.5 mg/mL, preferably 0.05 to 0.5mg/mL, for example 0.1 mg/mL. The concentration of carbetocin in theliquid (composition e.g. aqueous composition) may be, for example, 1mg/mL, 10 mg/mL, 50 mg/mL etc.

The concentration of the compound of formula (II) in the liquid(aqueous) composition may be from 0.01 to 4 mg/mL, for example 0.05 to 2mg/mL, more preferably 0.1 to 1.4 mg/mL, most preferably 0.2 to 0.7mg/mL.

The compositions of this aspect of the invention comprise ananti-oxidant. The anti-oxidant may be any anti-oxidant commonly used inthe art, for example any anti-oxidant approved for use as apharmaceutical excipient. For example, the anti-oxidant may bemethionine, EDTA, butylated hydroxy toluene, sodium metabisulfite etc.Preferably the anti-oxidant is present in an amount of 0.01% to 10%(w/v), for example 0.05% to 5% (w/v), most preferably 0.08% to 1% (w/v).Preferably the anti-oxidant is methionine, EDTA, or a combination ofmethionine and EDTA. In an example, the antioxidant is methionine and ispresent in an amount of 0.5% w/v. In an example, the antioxidant is EDTAand is present in an amount of 0.1% w/v.

The composition may further comprise an isotonicity agent. Isotonicityagents, for example mannitol or NaCl, are well known in the art.Preferably the isotonicity agent is present in an amount sufficient toprovide an isotonic composition (solution), for example in an amount of0.01% to 10% (w/v). Preferably the isotonicity agent is mannitol. If theisotonicity agent is mannitol it may be present in an amount of 0.5% to7.5% (w/v), more preferably 4.0% to 5.5% (w/v), for example 5.0% (w/v).If the isotonicity agent is mannitol it may be present in an amount of0.05% to 7.5% (w/v). If the isotonicity agent is NaCl it may be presentin an amount of 0.05% to 1.2% (w/v), more preferably 0.08% to 1% (w/v),for example 0.9% (w/v). The isotonicity agent may be present in anamount of 0.1 to 100 mg/mL, for example 0.5 to 7 mg/mL, for example 1 to5 mg/mL. For example, if the isotonicity agent is mannitol it may bepresent in an amount of 5 to 75 mg/mL, for example 40 to 55 mg/mL. Ifthe isotonicity agent is NaCl it may be present in an amount of 0.5 to12 mg/mL, for example 8 to 10 mg/mL.

The composition may be for any route of drug administration, e.g. oral,rectal, buccal, nasal, vaginal, transdermal (e.g. patch technology);parenteral, intravenous, intramuscular or subcutaneous injection;intracisternal, intravaginal, intraperitoneal, local (powders, ointmentsor drops) or as a buccal or nasal spray. Preferably the composition isfor nasal administration, e.g. is a nasal spray. As indicated above,compositions suitable for intranasal administration are expected toremain stable in solution at room temperature for an extended period oftime.

The composition may include an enhancer, an excipient which enhances theeffective dose (e.g. enhances the effective dose following nasaladministration). The enhancer may be any enhancer commonly used in theart, for example any enhancer approved for use as a pharmaceuticalexcipient. The enhancer may be, for example, methyl-β-cyclodextrin,Polysorbate 80, carboxymethylcellulose or hydroxypropylmethylcellulose.

It is preferred that the compositions of the invention do not include aquaternary amine compound, such as benzalkonium chloride. It ispreferred that the compositions of the invention do not include aparahydroxybenzoate preservative, or a combination ofparahydroxybenzoate preservative with a cosolvent. It is preferred thatthe compositions of the invention have a content of divalent metal ionsof less than 2 mM, for example 0.195 mM or less, for example 0.1 nM orless. It is preferred that compositions of the invention do not includea solubilizer. It is preferred that compositions of the invention do notinclude methyl-β-cyclodextrin.

According to the invention in a further aspect there is provided aliquid (e.g. aqueous) composition comprising: a pharmaceutically activecompound according to formula (II):

or (pharmaceutically acceptable) salt thereof; wherein the pH of thecomposition is from 5.0 to 6.0. The pH of the composition may be from5.0 to 5.9, for example from 5.1 to 5.9, for example 5.2 to 5.8. The pHof the composition may be from 5.1 to 6.0, for example from 5.2 to 6,for example from 5.26 to 6. The pH of the composition may be from 5.15to 5.75, for example from 5.2 to 5.65. The pH of the composition may befrom 5.26 to 5.8, for example from 5.26 to 5.75, for example from 5.26to 5.7, for example from 5.26 to 5.65, for example 5.4 to 5.65.Preferably, the liquid (e.g. aqueous) composition comprises apharmaceutically active compound according to formula (II):

The composition may be for any route of drug administration, e.g. oral,rectal, buccal, nasal, vaginal, transdermal (e.g. patch technology);parenteral, intravenous, intramuscular or subcutaneous injection;intracisternal, intravaginal, intraperitoneal, local (powders, ointmentsor drops) or as a buccal or nasal spray. Preferably the composition isfor nasal administration, e.g. is a nasal spray. As indicated above,compositions suitable for intranasal administration are expected toremain stable in solution at room temperature for an extended period oftime. Remarkably, the applicants have found that compositions of thecompound of formula (II) above may provide such stability at pH valueswhich are particularly suitable for nasal administration, withoutrequirement for inclusion of an anti-oxidant (see Example 7).

The composition may comprise a buffering agent, for example acetic acid,adipic acid, citric acid, maleic acid, succinic acid or (e.g. sodium)phosphate. The composition may include a single buffering agent. Thecomposition may include more than one buffering agent (e.g. may comprisecitric acid and (e.g. sodium) phosphate). The composition may comprise abuffer (solution), for example, a citrate buffer (solution), comprisingcitric acid and a citrate (e.g. sodium citrate); a succinate buffer(solution) comprising succinic acid and a succinate (e.g. sodiumsuccinate), an acetate buffer (solution) comprising acetic acid and anacetate (e.g. sodium acetate); a citrate/phosphate buffer (solution)comprising citric acid and phosphate; or a phosphate buffer (solution).

Preferably the pharmaceutical composition comprises a citrate/phosphatebuffer and the pH is from 5.1 to 6.0, for example from 5.2 to 6, forexample from 5.26 to 6.

Preferably the pharmaceutical composition comprises a succinate bufferor a citrate buffer and the pH is from 5.0 to 5.9, for example 5.0 to5.8, for example from 5 to 5.7.

The concentration of the compound of formula (II) in the liquid(aqueous) composition may be from 0.01 to 4 mg/mL, for example 0.05 to 2mg/mL, more preferably 0.1 to 1.4 mg/mL, most preferably 0.2 to 0.7mg/mL.

The compositions of this aspect of the invention further comprise ananti-oxidant. The anti-oxidant may be any anti-oxidant commonly used inthe art, for example any anti-oxidant approved for use as apharmaceutical excipient. For example, the anti-oxidant may bemethionine, EDTA, butylated hydroxy toluene, sodium metabisulfite etc.Preferably the anti-oxidant is present in an amount of 0.01% to 10%(w/v), for example 0.05% to 5% (w/v), most preferably 0.08% to 1% (w/v).Preferably the anti-oxidant is methionine, EDTA, or a combination ofmethionine and EDTA. In an example, the antioxidant is methionine and ispresent in an amount of 0.5% w/v. In an example, the antioxidant is EDTAand is present in an amount of 0.1% w/v.

The composition may further comprise an isotonicity agent. Isotonicityagents, for example mannitol or NaCl, are well known in the art.Preferably the isotonicity agent is present in an amount sufficient toprovide an isotonic composition (solution), for example in an amount of0.01% to 10% (w/v). Preferably the isotonicity agent is mannitol. If theisotonicity agent is mannitol it may be present in an amount of 0.5% to7.5% (w/v), more preferably 4.0% to 5.5% (w/v), for example 5.0% (w/v).If the isotonicity agent is mannitol it may be present in an amount of0.05% to 7.5% (w/v). If the isotonicity agent is NaCl it may be presentin an amount of 0.05% to 1.2% (w/v), more preferably 0.08% to 1% (w/v),for example 0.9% (w/v). The isotonicity agent may be present in anamount of 0.1 to 100 mg/mL, for example 0.5 to 7 mg/mL, for example 1 to5 mg/mL. For example, if the isotonicity agent is mannitol it may bepresent in an amount of 5 to 75 mg/mL, for example 40 to 55 mg/mL. Ifthe isotonicity agent is NaCl it may be present in an amount of 0.5 to12 mg/mL, for example 8 to 10 mg/mL.

It is preferred that the compositions of the invention do not include aquaternary amine compound, such as benzalkonium chloride. It ispreferred that the compositions of the invention do not include aparahydroxybenzoate preservative, or a combination ofparahydroxybenzoate preservative with a cosolvent. It is preferred thatthe compositions of the invention have a content of divalent metal ionsof less than 2 mM, for example 0.195 mM or less, for example 0.1 nM orless. It is preferred that compositions of the invention do not includea solubilizer. It is preferred that compositions of the invention do notinclude methyl-β-cyclodextrin.

According to the present invention in a further aspect, there isprovided a method of treatment or prevention of uterine atony [forexample, treatment or prevention of uterine atony following vaginaldelivery of the infant, treatment or prevention of uterine atonyfollowing delivery of the infant by Caesarean section, for exampledelivery of the infant by Caesarean section under epidural or spinalanaesthesia, or treatment or prevention of uterine atony in a patientwho is at risk of developing PPH], or a method of treatment orprevention of bleeding (e.g. excessive bleeding) following vaginaldelivery (of the infant), comprising: administration to a patient inneed thereof a liquid (e.g. aqueous) pharmaceutical compositioncomprising carbetocin or a pharmaceutically active salt thereof; whereinthe pH of the composition is from 5.0 to 6.0. The pH of the compositionmay be from 5.1 to 6.0, for example from 5.2 to 6, for example from 5.26to 6. The pH of the composition may be from 5.15 to 5.75, for examplefrom 5.2 to 5.65. The pH of the composition may be from 5.26 to 5.8, forexample from 5.26 to 5.75, for example from 5.26 to 5.7, for examplefrom 5.26 to 5.65, for example 5.4 to 5.65.

According to the present invention in a further aspect, there isprovided a kit of parts comprising: a liquid (e.g. aqueous)pharmaceutical composition comprising carbetocin or a pharmaceuticallyactive salt thereof wherein the pH of the composition is from 5.0 to6.0; and a container [e.g. ampoule, vial, pre-filled syringe, injectiondevice (e.g. single use injection device such as that sold under themark Uniject by Becton Dickinson), injection cartridge, ampoule,multi-dose pen] for the composition, optionally with separate injectionmeans (e.g. if required for administration), optionally withinstructions for administration of the composition. The pH of thecomposition may be from 5.1 to 6.0, for example from 5.2 to 6, forexample from 5.26 to 6. The pH of the composition may be from 5.15 to5.75, for example from 5.2 to 5.65. The pH of the composition may befrom 5.26 to 5.8, for example from 5.26 to 5.75, for example from 5.26to 5.7, for example from 5.26 to 5.65, for example 5.4 to 5.65.

According to the present invention in a further aspect, there isprovided a kit of parts comprising: a liquid (e.g. aqueous)pharmaceutical composition comprising a pharmaceutically active compound(e.g. carbetocin) or salt thereof and optionally an anti-oxidant,wherein the pH of the composition is from 5.0 to 6.0; and a container(e.g. vial, pre-filled syringe, injection device [e.g. single usepre-filled injection device such as that sold under the mark Uniject byBecton Dickinson), injection cartridge, ampoule, multi-dose pen] for thecomposition, optionally with separate injection means (e.g. if requiredfor administration), optionally with instructions for administration ofthe composition. The pH of the composition may be from 5.1 to 6.0, forexample from 5.2 to 6, for example from 5.26 to 6. The pH of thecomposition may be from 5.15 to 5.75, for example from 5.2 to 5.65. ThepH of the composition may be from 5.26 to 5.8, for example from 5.26 to5.75, for example from 5.26 to 5.7, for example from 5.26 to 5.65, forexample 5.4 to 5.65.

According to the present invention in a further aspect, there isprovided a method of treatment or prevention of compromised lactationconditions, labour induction impairment, uterine atony conditions,excessive bleeding, inflammation, pain, abdominal pain, back pain, maleand female sexual dysfunction, irritable bowel syndrome (IBS),constipation, gastrointestinal obstruction, autism, stress, anxiety,depression, anxiety disorder, surgical blood loss, post-partumhaemorrhage, wound healing, infection, mastitis, placenta deliveryimpairment, osteoporosis, and a method for the diagnosis of cancer andplacental insufficiency, comprising: administration to a patient in needthereof a liquid (e.g. aqueous) pharmaceutical composition comprising apharmaceutical compound according to formula (I) or (II) as definedabove or a pharmaceutically active salt thereof; wherein the pH of thecomposition is from 5.0 to 6.0. According to the present invention in astill further aspect, there is provided a liquid (e.g. aqueous)pharmaceutical composition comprising a pharmaceutical compoundaccording to formula (I) or (II) as defined above or a pharmaceuticallyactive salt thereof; wherein the pH of the composition is from 5.0 to6.0; for use in (or in the manufacture of a medicament for) thetreatment or prevention of compromised lactation conditions, labourinduction impairment, uterine atony conditions, excessive bleeding,inflammation, pain, abdominal pain, back pain, male and female sexualdysfunction, irritable bowel syndrome (IBS), constipation,gastrointestinal obstruction, autism, stress, anxiety, depression,anxiety disorder, surgical blood loss, post-partum haemorrhage, woundhealing, infection, mastitis, placenta delivery impairment,osteoporosis, or for use in (or in the manufacture of a substance for)the diagnosis of cancer or placental insufficiency. The pH of thecomposition may be from 5.1 to 6.0, for example from 5.2 to 6, forexample from 5.26 to 6. The pH of the composition may be from 5.0 to5.9, for example from 5.1 to 5.9, for example 5.2 to 5.8. The pH of thecomposition may be from 5.15 to 5.75, for example from 5.2 to 5.65. ThepH of the composition may be from 5.26 to 5.8, for example from 5.26 to5.75, for example from 5.26 to 5.7, for example from 5.26 to 5.65, forexample 5.4 to 5.65.

According to the present invention in a further aspect, there isprovided a kit of parts comprising: a liquid (e.g. aqueous)pharmaceutical composition comprising carbetocin or a pharmaceuticallyactive compound according to formula (II):

or a pharmaceutically active salt thereof wherein the pH of thecomposition is from 5.0 to 6.0; and a container [e.g. ampoule, vial,pre-filled syringe, injection device (e.g. single use injection devicesuch as that sold under the mark Uniject by Becton Dickinson), injectioncartridge, ampoule, multi-dose pen] for the composition, optionally withseparate injection means (e.g. if required for administration),optionally with instructions for administration of the composition. ThepH of the composition may be from 5.0 to 5.9, for example from 5.1 to5.9, for example 5.2 to 5.8. The pH of the composition may be from 5.1to 6.0, for example from 5.2 to 6, for example from 5.26 to 6. The pH ofthe composition may be from 5.15 to 5.75, for example from 5.2 to 5.65.The pH of the composition may be from 5.26 to 5.8, for example from 5.26to 5.75, for example from 5.26 to 5.7, for example from 5.26 to 5.65,for example 5.4 to 5.65.

According to the present invention in a further aspect, there isprovided a kit of parts comprising: a liquid (e.g. aqueous)pharmaceutical composition comprising a pharmaceutically active compound(e.g. carbetocin, a compound of formula (I) or (II) above) or saltthereof and optionally an anti-oxidant, wherein the pH of thecomposition is from 5.0 to 6.0; and a container (e.g. vial, pre-filledsyringe, injection device [e.g. single use pre-filled injection devicesuch as that sold under the mark Uniject by Becton Dickinson), injectioncartridge, ampoule, multi-dose pen] for the composition, optionally withseparate injection means (e.g. if required for administration),optionally with instructions for administration of the composition. ThepH of the composition may be from 5.1 to 6.0, for example from 5.2 to 6,for example from 5.26 to 6. The pH of the composition may be from 5.0 to5.9, for example from 5.1 to 5.9, for example 5.2 to 5.8. The pH of thecomposition may be from 5.15 to 5.75, for example from 5.2 to 5.65. ThepH of the composition may be from 5.26 to 5.8, for example from 5.26 to5.75, for example from 5.26 to 5.7, for example from 5.26 to 5.65, forexample 5.4 to 5.65.

DETAILED DESCRIPTION OF THE INVENTION

The present invention will now be illustrated with reference to theattached drawings in which:

FIG. 1 shows a UPLC chromatogram of an impurity mix of carbetocin anddegradation products;

FIG. 1 a shows the chemical formulae of carbetocin and degradationproducts;

FIG. 2 shows the content of degradation (hydrolysis) product[Gly⁹OH]carbetocin (see FIG. 1 a ) in the antioxidant study samples as afunction of time (constant pH);

FIG. 3 shows the content of degradation (oxidation) product sulfoxide IIcarbetocin (see FIG. 1 a ) in the antioxidant study samples as afunction of time (constant pH);

FIG. 4 shows individual degradation products at different pH (pH study,constant antioxidant);

FIG. 5 shows the sum of degradation products at different pH (pH study,constant antioxidant),

FIG. 6 shows the Stability of FE 202767 in selected buffers; and

FIG. 7 shows the sum of impurities of various carbetocin formulationsafter 12 months at 40° C. and 75% relative humidity (R.H.), as describedin Experiment 3A.

ANALYTICAL METHOD

This is the analytical method for the carbetocin examples (Examples 1 to6), below.

All solutions were analysed on a Waters Acquity UPLC (Ultra-highPressure Liquid chromatography) system using isocratic conditions. TheMobile phase was 20% acetonitrile (JT Baker, Ultra Gradient Grade) in 5mM unbuffered ammonium acetate (Fluka, Ultra≥99.0%). The column was aWaters Acquity UPLC BEH Shield RP18, 2.1*100 mm, 1.7 μm (Flow: 0.5ml/min, Column Temp: 50° C.). The injection volume was 20 μl. Detectionwas performed by UV at 220 nm. The different impurities were evaluatedas area % of total area.

FIG. 1 shows a chromatogram of an impurity mix of carbetocin and itsdegradation products. The solution contained, carbetocin, the hydrolysisproducts [Gly⁹OH], [Asp⁵] and [Glu⁴]carbetocin, the oxidation productssulfoxide I and sulfoxide II-carbetocin, the alkaline degradationproducts [βAsp⁵] and [D-Asn⁵]carbetocin and the synthesis relatedimpurity [D-Cys⁶]carbetocin. The chemical formulae of carbetocin and thedegradation products (hydrolysis products, oxidation products andalkaline degradation products) are shown in FIG. 1 a . The“[Glu⁴]carbetocin” type nomenclature is well known in the art. Theresolutions between all peaks were ≥2.0.

Example 1: Formulation Antioxidant Study (Constant pH)

5.0 grams of D(−)-Mannitol (Ph Eur, Prolabo) was dissolved in 1000 ml ofmilliQ-water. This solution was adjusted with acetic acid (Ph.Eur.,Merck) to pH 5.2. This solution was then divided into four 200 mlaliquots. To aliquot 1, 0.2 gram of EDTA disodium, dihydrate (Fluke) wasadded and dissolved. To aliquot 2, 1.0 gram of L-methionine (Sigma,non-animal source) was added and dissolved. To aliquot 3, 0.2 gram ofEDTA disodium, dihydrate and 1.0 g of L-methionine was added anddissolved. Nothing was added to aliquot 4. The pH of aliquots 1-3 wasadjusted with acetic acid to pH 5.2±0.1. 1 mg of carbetocin (PolypeptideLaboratories) was transferred to four 10 ml volumetric flasks. Aliquots1-4 were used to dissolve the substance and for dilution to volume (0.1mg/ml carbetocin). The solutions were transferred to 25 ml blue capflasks, and placed in a cabinet at 40° C. and 75% RH. A sample of thecurrent PABAL® formulation, pH 3.9 (measured), was placed in the samecabinet for comparison.

The solutions were analysed after 2, 6, 12, 22 and 33 weeks at 40° C.The largest impurities of this study was found to be the hydrolysisproduct [Gly⁹OH]carbetocin and the oxidation product [sulfoxideII]carbetocin. The pH in this study (pH 5.2) was not high enough tostart any alkaline degradation of carbetocin. The content % (w/w) of themajor impurity formed by hydrolysis, [Gly⁹OH]carbetocin, and byoxidation, sulfoxide II-carbetocin, are shown in FIGS. 2 and 3 . Theproduct specification allowed for each impurity is also plotted in eachfigure as a reference. Thus it can be seen from FIG. 2 that if theconcentration of [Gly⁹OH] carbetocin increases above 1.5% the sample is“out of specification”, that is the sample has degraded such that is nolonger suitable for administration.

As shown in FIG. 2 , the antioxidant study (constant pH) showed that theformation of hydrolysis products, mainly [Gly⁹OH]carbetocin, was veryfast in the current PABAL formulation (pH 3.9). With regard to thecontent of [Gly⁹OH]carbetocin the current formulation was quicklyout-of-specification (>1.5%), after 6 weeks at 40° C. All formulationsat pH 5.2 were well below the specification limit after 33 weeks at 40°C. (0.4-0.6%), indicating that these formulations are stable for atleast 6 months at 40° C. and 75% RH, which is generally accepted toindicate a likely stability for at least 24 months at 25° C. and 60% RH(i.e. a stable RT formulation) The results applied for all threehydrolysis products. In FIG. 3 , the addition of antioxidant was shownto be very effective in slowing down the oxidation of carbetocin,despite the increased pH of the formulations. The formulation at pH 5.2that did not contain any additives was out-of-specification (>0.8%) withregard to the content of sulfoxide II-carbetocin after approx 20 weeksat 40° C. The formulations containing methionine or EDTA were all wellbelow the specification limit after 33 weeks (0.2-0.4%). The formulationcontaining a combination of EDTA and methionine did not show anyincrease of oxidation products at all, compared to the levels found inthe substance batch. Due to the low pH, the current formulation was notprone to degradation by oxidation (pH 3.9). The results are shown innumerical form in the following table (Table 1).

TABLE 1 Individual and sum of degradation products (%) after 33 weeks at40° C. (constant pH). Sulfoxide Sulfoxide Sum of Formulation Gly⁹OH Asp⁵Glu⁴ I II βAsp5 D-Asn⁵ impurities Current formulation 6.43 1.15 5.410.42 0.38 0.13 0.15 16.4 Mannitol pH 5.2 0.53 0.14 0.42 0.51 0.93 0.130.12 3.5 Mannitol pH 0.63 0.20 0.51 0.14 0.25 0.15 0.16 3.2 5.2 +methionine Mannitol pH 0.52 0.15 0.38 0.31 0.39 0.15 0.10 2.9 5.2 + EDTAMannitol pH 0.44 0.13 0.35 0.10 0.16 0.19 0.17 2.4 5.2 + methionine +EDTA

Table 1 includes the sum of degradation products for all samples, andafter 33 weeks the effect of EDTA is clearer. Further, the samplecontaining both methionine and EDTA is clearly better than the others.The evidence points to a linear degradation: assuming this is indeed thecase, the mannitol pH 5.2+methionine+EDTA sample is likely to be inspecification—i.e., suitable for use—for a remarkable 86 weeks at 40° C.This is based, as is well known in the art, on a linear extrapolation ofthe increase in impurity over time to determine when the amount ofimpurity would be sufficiently high for the formulation to be “out ofspecification”.

Example 2: Formulation pH Study (Constant Antioxidant)

1.2 grams of succinic acid (Sigma-Aldrich, 99%) and 1.0 g ofL-methionine (Sigma, non-animal source) were dissolved in 1000 ml ofmilliQ water (10 mM). This solution was adjusted, in aliquots, withdiluted NaOH (Ph.Eur., Merck) to pH 4.0, 4.5, 5.2, 5.65, 6.1, 6.5 and7.0. 55 mg of carbetocin (Polypeptide Laboratories) was dissolved in 50ml of milliQ water (1.1 mg/ml). 1.0 ml of the carbetocin solution (1.1mg/ml) was mixed with 10 ml of each buffer (0.1 mg/ml carbetocin). 0.55g of mannitol (5%) was added to each solution and dissolved. Thesolutions were transferred to 15 ml glass vials with screw lid andplaced in a 40° C. cabinet at 75% RH.

The solutions were analysed after 12 and 52 weeks at 40° C. The contentof the individual degradation products and the sum of degradationproducts is presented in Tables 2a and 2b and FIGS. 4 and 5 .

TABLE 2a Individual and sum of degradation products (%) at different pHafter 12 weeks at 40° C. (pH study, constant antioxidant). Oxidationproducts Alkaline impurities Sample Hydrolysis products SulfoxideSulfoxide Unknown pH [Gly⁹OH] [Asp⁵] [Glu⁴] I II [D-Asn⁵] [β Asp⁵] 2.0min Sum 4.0 2.22 0.50 1.78 N.D N.D N.D. N.D. N.D. 4.50 4.5 0.84 0.190.67 N.D N.D N.D. N.D. N.D. 1.70 5.2 0.23 0.10 0.27 N.D N.D 0.06 0.050.09 0.80 5.65 0.15 0.04 0.12 N.D 0.03 0.12 0.08 0.16 0.70 6.1 0.14 0.090.13 0.03 0.03 0.30 0.22 0.32 1.26 6.5 0.25 0.13 0.12 0.03 0.03 0.540.30 0.48 1.88 7.0 0.52 0.20 0.17 0.02 0.02 1.27 0.47 0.62 3.29

TABLE 2b Individual and sum of degradation products (%) at different pHafter 52 weeks at 40° C. (pH study, constant antioxidant). Oxidationproducts Alkaline impurities Sample Hydrolysis products SulfoxideSulfoxide Unknown pH [Gly⁹OH] [Asp⁵] [Glu⁴] I II [D-Asn⁵] [β Asp⁵] 2.0min Sum 4.0 7.55 1.09 6.23 0.04 0.07 0.00 0.19 0.94 19.3 4.5 2.92 0.572.49 0.04 0.05 0.00 0.19 0.40 7.8 5.2 0.78 0.26 0.72 0.05 0.14 0.26 0.250.22 3.6 5.65 0.48 0.22 0.52 0.02 0.02 0.45 0.40 0.40 3.8 6.1 0.56 0.350.34 0.08 0.11 1.06 0.73 0.77 5.9 6.5 0.88 0.49 0.32 0.03 0.06 2.30 1.251.31 8.8 7.0 1.53 0.71 0.43 0.05 0.04 4.10 1.68 1.65 12.9

As discussed below with reference to Example 3a, the specification limitfor sum of impurities (for the current PABAL® formulation) is ≤5%. Ascan be seen from Table 2b (“Sum”), the Samples at pH 5.2 and 5.65(examples of the invention) are still within specification after 52weeks (1 year) at 40° C., while all other samples are out ofspecification after 52 weeks (1 year) at 40° C.

The results of the pH study (FIGS. 4, 5 ) confirmed the observations ofthe antioxidant study. The formation of hydrolysis products ([Gly⁹OH],[Asp⁵] and [Glu⁴]carbetocin) was effectively reduced by an increase inpH from pH 4.0 to about pH 5.65. At higher pH values (pH 6.1-7.0) thecontent of hydrolysis products was again increased. The effectiveness ofthe antioxidant at about 1 mg/ml concentration was also confirmed (inthe antioxidant study the concentration of antioxidant was 5 mg/ml).However, if oxidation is limited in the drug or drug solution (e.g. ifthe drug or drug solution is not prone to oxidation), the amount ofantioxidant may be reduced, or use of antioxidant may not be necessary.Due to the antioxidant, the oxidation of carbetocin was negligible,regardless of pH. The upper pH limit of an optimised formulation wasinstead limited by the alkaline degradation of carbetocin. The mainimpurity by alkaline degradation was [D-Asn⁵]carbetocin, which wasrapidly increased at pH values above pH 6.1. The formation of two otherminor impurities at high pH was also observed, [βAsp⁶]carbetocin and oneunknown impurity eluting early in the chromatogram (tR: 2.0 min).

The U-shape of the pH vs. sum of degradation products curve illustratesthe stability plateau of carbetocin at pH 5.0-6.0. At pH 5.2 the sum ofdegradation products was found to be only 16% of the sum of degradationproducts at pH 4.0 (current formulation). The optimal pH was found to besomewhere between pH 5.1 to 6, for example between around pH 5.2 and5.65.

Examples 1 and 2 give a very strong indication that formulations of theinvention are room temperature stable for up to two years.

Example 3: Formulation Study of Isotonicity Agents, NaCl Vs. Mannitol,at 30° C., 40° C.

4.22 grams of citric acid monohydrate (Merck, pro analysi) was dissolvedin 2000 ml of milliQ water (20 mM). This solution was divided into ten200 mi aliquots. 1.8 g of sodium chloride (Merck, pro analysi) was addedto five of the flasks, to the other five flasks 10 g of mannitol (VWR,Ph Eur) was added. According to an experimental design, 0.2, 0.6 or 1.0g of L-methionine (Sigma, non-animal source) was added and the pH wasadjusted with 1% NaOH (Merck, pro analysi) to pH 5.2, 5.65 or 6.1, seeTable 3a and 3b. 2 mg of Carbetocin (Polypeptide Laboratories) wastransferred to twelve 20 ml volumetric flasks and the substance wasdissolved in each buffer (0.1 mg/ml carbetocin). The samples containing3 mg/ml methionine were prepared in duplicate, see Table 3a and 3b.

Two mL of each solution was transferred to LC-vials and placed in a 30°C./75% RH cabinet. The remaining solutions were transferred to 25 mlblue cap flasks and placed in a 40° C./75% R.H. cabinet. The level ofimpurities after 25 weeks in 30′C/75% R.H are shown in the followingTables 3a and 3b.

TABLE 3a Sulfoxide Sulfoxide Unknown Formulation Gly⁹OH Asp⁵ Glu⁴ I II 2min βAsp⁵ D-Asn⁵ D-Cys⁶ Sum ** Mannitol, pH 0.18 0.04 0.18 0.04 0.03N.D. N.D. N.D. 0.09 0.80 5.2, 1 mg/mL methionine Mannitol, pH 0.07 0.030.07 0.03 N.D. 0.17 0.15 0.01 0.12 0.95 6.1, 1 mg/mL methionineMannitol, pH 0.10 0.03 0.08 0.04 0.05 0.09 0.07 0.05 0.12 0.78 5.65, 3mg/mL methionine, sample 1 Mannitol, pH 0.09 0.02 0.11 0.03 N.D. 0.080.04 N.D. 0.11 0.63 5.65, 3 mg/mL methionine, sample 2 Mannitol, pH 0.170.03 0.20 0.02 N.D. N.D. 0.04 N.D. 0.11 0.75 5.2, 5 mg/mL methionineMannitol, pH 0.10 0.05 0.05 0.04 N.D. 0.23 0.18 0.11 0.12 1.11 6.1, 5mg/mL methionine Years to OOS* 8 12 7 “infinity” “infinity” 6 12“infinity” N/A 4.2 for 3 mg/mL, pH 5.65 sample 2. *Out of specification** of degradation products

TABLE 3b (NaCl) Sulfoxide Sulfoxide Unknown Beta Formulation Gly⁹OH Asp⁵Glu⁴ I II 2 min Asp⁵ D-Asn⁵ D-Cys⁸ Sum ** NaCl, pH 5.2, 0.21 0.06 0.150.02 0.03 ND 0.02 0.12 0.82 1 mg/mL methionine NaCl, pH 6.1, 0.08 0.050.04 0.04 0.04 0.2  0.12 0.12 0.11 0.99 1 mg/mL methionine NaCl, pH5.65, 0.09 0.03 0.07 0.01 ND 0.12 0.04 0.05 0.11 0.74 3 mg/mL methioninesample 1 NaCl, pH 5.65, 0.09 0.04 0.08 0.02 0.03 0.10 0.05 0.14 0.110.78 3 mg/mL methionine sample 2 NaCl, pH 5.2, 0.17 0.04 0.15 0.02 0.04ND 0.05 0.14 0.86 5 mg/mL methionine NaCl, pH 6.1, 0.09 0.04 0.06 0.03ND 0.23 0.17 0.14 0.11 1.11 5 mg/mL methionine ** of degradationproducts

Tables 3a and Tables 3b show that there is very little degradation inall samples. This level of degradation corresponds to that seen after 6weeks at 40° C.

The results indicate that the best samples are likely to be stable for 5years at 30° C. As seen in Table 3a, rows 5 and 8, results for themethionine 3 mg/mL, pH 5.65 sample 2 indicate that this sample wouldremain in specification for more than 4 years at 30° C. and 75% RH. Thisis based, as is well known in the art, on a linear extrapolation of theincrease in impurity over time to determine when the amount of impuritywould be sufficiently high for the formulation to be “out ofspecification” (OOS). It was also found that the optimum pH at 30° C. ishigher than at 40° C. (results not shown). The differences are small butpH 5.65 is slightly superior to pH 5.2 at 30° C. (vice versa at 40° C.).These results indicate there is a good margin for obtaining a climatezone III/IV stable formulation.

The applicants found that increase in methionine leads to moredegradation, mainly by increase of [BetaAsp5]carbetocin. A concentrationof about 1 mg/ml appears to be sufficient to provide effectivestabilisation without significant degradation.

Experiment 3a—the Stability of Carbetocin at Different pH and UsingDifferent Antioxidants

This study was designed to give a broader picture of the stability ofcarbetocin at different pH and using different antioxidants.

1.2 grams of succinic acid (Sigma-Aldrich, ≥99%) was dissolved in 1000ml of milliQ water (10 mM). This solution was adjusted, in aliquots,with diluted NaOH (Ph.Eur., Merck) to pH 4.0, 4.5, 5.2, 5.65, 6.1, 6.5and 7.0. 55 mg of carbetocin (Polypeptide Laboratories, Strasbourg) wasdissolved in 50 ml of milliQ water (1.1 mg/ml). 1.0 ml of the carbetocinsolution (1.1 mg/ml) was mixed with 10 ml of each buffer (0.1 mg/mlcarbetocin). 0.55 g of mannitol (5%) was added to each solution anddissolved. The solutions were transferred to 15 ml glass vials withscrew lid and placed in the 40° C./75% R.H. cabinet.

The same procedure was repeated; with the exception that 1.0 g ofL-methionine (Sigma, non-animal source) was added to the 1000 ml ofmilliQ water, giving duplicate samples containing 1 mg/ml methionine atall pH-levels. The solutions were transferred to 15 ml glass vials withscrew lid and placed in the 40° C./75% R.H. cabinet.

The buffers at pH 5.65, 6.1 and 6.5 were also divided into aliquots towhich EDTA disodium, dihydrate (Fluka) was added. These samples werestored at 40° C./75% for 12 months before analysis.

The sum of impurities after 12 months at 40° C./75% R.H are shown inFIG. 7 . The Figure also shows the “spec limit”, above which the sum ofimpurities is such that the formulation is out of specification.

All formulations at pH 5.2 and pH 5.65 were within specification after12 months at 40° C./75% R.H.

The positive effect of methionine was visible also in this study. Allsamples containing methionine showed very low amounts of oxidationproducts, regardless of composition and pH. This points to inclusion ofmethionine in a robust formulation, where (for example) metal ioncontent of the active ingredient carbetocin, which can vary withproduction batch and which, if high, may lead to increased oxidation,will not be a controlled parameter.

The most stable formulation was the formulation at pH 5.2 containing 1mg/ml of methionine (results not shown). The parameter that was closestto the specification limit after 12 months at 40° C./75% R.H. was thesum of impurities (FIG. 7 ). The specification limit for sum ofimpurities (for the current PABAL® formulation) is ≤5% (i.e. 5.5%). Itcan be assumed that the degradation is linear over time, and it cantherefore be calculated that the formulation at pH 5.2 containing 1mg/ml of methionine would be out-of-specification after approximately 80weeks at 40° C./75% R.H, based on the specification for the currentPABAL® formulation.

A commonly used guide, supported by the Arrhenius equation, is that therate of most chemical reactions doubles for every 10° C. increase oftemperature. If we apply this relationship to the formulation at pH 5.2containing 1 mg/ml of methionine, the estimated shelf-life of a newformulation will be 160 weeks at 30° C., i.e. slightly more than 3years, again based on the specification for the current PABAL®formulation. This is likely to be an underestimation, since the reported“sum of impurities” in this experiment included every peak on thebaseline, including synthesis related impurities and peaks below thereporting limit (<0.05%). The synthesis related impurities consistmainly of [DCys⁶] and [desGln⁴]carbetocin, which do not increase duringstorage. The substance batch contained 0.9% impurities according to thesupplier. Thus, it is likely that a shelf-life of more than 3 years at30° C./75% R.H. would be achieved for this formulation.

Example 4—Formulation in Succinate Buffer

The following preparation and decanting was performed in apharmaceutical room under germ free conditions. 47 grams of mannitol,1.2 grams of succinic acid buffering agent and 1.0 g of L-methionine wasdissolved in about 900 ml of milliQ water (10 mM). The pH of thesolution was adjusted with 5M NaOH to pH 5.4. The solution wastransferred to a 1000 ml volumetric flask and diluted to volume withWFI.

50 mg of carbetocin (Polypeptide Laboratories) was transferred to a 500ml volumetric flask and dissolved and diluted to volume with themannitol/succinic acid/methionine buffer pH 5.4. The solution wasfiltered through a 0.22 μm filter and filled in glass vials with rubberstoppers (1.1 ml per vial). Each vial included an aqueous compositioncomprising carbetocin (0.1 mg/mL), and the pH of the composition was 5.4(i.e. from 5.0 to 6.0). The aqueous composition also included succinatebuffer (succinic acid buffering agent), methionine (anti-oxidant) andmannitol (isotonic agent). In a further Example (Example 4A, not shown)a solution was made up exactly as Example 4 and EDTA (0.1% w/v) added.The osmolality of the solutions in Example 4 and 4A was found to be300±20 mOsmol/kg.

The formulation of Example 4 (and that of Example 4A) is suitable forinjection to a patient with uterine atony.

Example 5—Formulation in Succinate Buffer

The following preparation and decanting was performed in apharmaceutical room under germ free conditions. 1.2 grams of succinicacid buffering agent (Sigma-Aldrich, 99%) and 1.0 g of L-methionine(Sigma, non-animal source) were dissolved in 1000 ml of milliQ water (10mM) to provide a succinate buffer of pH 5.4, the pH being adjusted tothis value with NaOH solution.

0.55 g of mannitol (5%) was dissolved in 10 ml of succinate buffer.Methionine 0.5% (w/v) was added to the solution and dissolved.Carbetocin (Polypeptide Laboratories) was dissolved in the solution sothe concentration of carbetocin was 0.1 mg/mL, and the pH adjusted to5.4 using NaOH solution. The solution was divided into 1 mL quantitiesand sealed in ampoules. Each ampoule included an aqueous compositioncomprising carbetocin (0.1 mg/mL), and the pH of the composition was 5.4(i.e. from 5.0 to 6.0). The aqueous composition also included succinatebuffer (succinic acid buffering agent), methionine (anti-oxidant) andmannitol (isotonic agent). It will be appreciated that the compositionmay be made with water for injection (WFI). The formulation of Example 5is suitable for injection to a patient with uterine atony.

Example 6—Formulation with Citrate/Phosphate Buffer

The formulation set out in the following table was made up by similarmethods to those set out in Examples 4 and 5 above.

TABLE 4 Component Amount per mL Function Carbetocin 10 mg Activeingredient Sodium phosphate dibasic 3.24 mg Buffering agent dihdrateCitric acid monohydrate 1.43 mg Buffering agent NaCl 7.5 mg Isotonicityagent HCl q.s. adjust to pH 5.5 pH adjustment NaOH q.s. adjust to pH 5.5pH adjustment Water for Injection Adjust to 1 mL SolventThe composition is suitable for nasal administration.Optionally, an antioxidant (e.g. methionine at a concentration of 1.0mg/mL may be included in the formulation). The anti-oxidant may be anyanti-oxidant commonly used in the art.Optionally, the composition may include an enhancer. The enhancer may beany enhancer commonly used in the art, for example any enhancer approvedfor use as a pharmaceutical excipient. The enhancer may be, for example,methyl-β-cyclodextrin, Polysorbate 80, carboxymethylcellulose orhydroxypropylmethylcellulose.

Example 7—the Stability of FE 202767 in Citrate and Citrate-PhosphateBuffers (pH 5.0, 5.5, and 6.0) at 40° C. for a Six Month Period.Materials and Methods

FE 202767 (Ferring) was synthesised by the method set out inWO2009/122285. FE 202767 was dissolved at a concentration of 0.2 mg/mlin either 25 mM citrate buffer (isotonic to saline) or 25 mMcitrate-phosphate buffer (isotonic with saline) at varying pH (pH 5.0,5.5, 6.0), by methods known in the art. The solutions were incubated at40° C. for 176 days, with samples taken at day 0, 15, 30, 84, and 176.Samples were evaluated by HPLC to determine the amount of intact peptideremaining at the various time points, by methods well known in the art,comparing the % Area. of the intact peptide peak on the sampling day vs.% Area on Day 0.The HPLC method used an Agilent 1200 instrument. The mobile phases whereHPLC Buffers A (A=0.01% TFA in water) and B (B=0.01% TFA in [70% v/vacetonitrile and 30% v/v water]) with the gradient 15% B for 1 min, then15 to 95% B in 30 min, then 95 to 100% B in 3 min, then 100% B for 5 minand 100% B to 15% B in 1 min at flow rate 0.3 mL/min. The PhenomenexMAX-RP C18, 2.0×150 mm, 4 μm, 80 Å column was at temperature 40 with UVdetection at 210 nm. The injection volume was 10 μL.The results are shown in the following Table 5, and on the attached FIG.6 . In Table 5 and FIG. 6 , CP50 is citrate phosphate buffer at pH 5.0;CP55 is citrate phosphate buffer at pH 5.5; and CP60 is citratephosphate buffer at pH 6.0; CT50 is citrate phosphate buffer at pH 5.0;CT55 is citrate phosphate buffer at pH 5.5; and CT60 is citratephosphate buffer at pH 6.0.

TABLE 5 % Intact Peptide Remaining (normalised to day 0) Day CP50 CP55CP60 CT50 CT55 CT60 0 100.00 100.00 100.00 100.00 100.00  100.00 6 99.9799.83 99.65 99.66 99.75 100.00 15 99.66 99.31 99.54 99.51 99.71 99.40 3099.47 99.30 99.26 99.29 n.a. 99.30 84 98.38 97.44 97.78 98.05 98.6197.94 176 96.98 95.60 95.48 95.19 97.34 73.36 Notes: % Intact PeptideRemaining expressed relative to % Area on Day 0. n.a. = data pointexcluded due to aberrant peak in HPLC chromatogram. CP =citrate-phosphate buffer; CT = citrate buffer.

Conclusion

FE 202767 showed good stability in citrate-phosphate buffers in the pHrange tested (pH 5.0, 5.5, and 6.0), with >95% remaining after 176 daysin each condition. It was also very stable (>95% remaining) in citratebuffer at pH 5.0 and 5.5; however, there was significant degradationafter 176 days in pH 6.0 citrate buffer.In general, a formulation suitable for nasal administration is expectedto be of pH between 5.0 and 6.0, include the minimum number of reagents(e.g. no anti-oxidant). It is also preferred that the formulation isroom temperature stable. Example 7 demonstrates that formulations alongthe lines above may be suitable for nasal administration, because theyhave appropriate pH and are room temperature stable without requirementfor anti-oxidant or other additives that might adversely affect thenasal mucosa.

Example 8—the Stability of FE 202767 in Various Buffers at 40° C. forOne and Three Months Materials and Methods

The method was similar to Example 7. FE 202767 (Ferring) was synthesisedby the method set out in WO2009/122285. The FE 202767 was dissolved at aconcentration of 0.2 mg/ml in either 25 mM citrate buffer (citricacid/Na citrate), 10 mM acetate buffer (acetic acetate/Na acetate) or 10mM succinate buffer (1 mM succinic acid+NaOH to relevant pH) at varyingpH (pH 5.0, 5.2, 5.5, 5.65, 5.8, 6.0), by methods known in the art. Asset out in the table below, the various samples also includedisotonicity agent (NaCl, 7 mg/mL or mannitol 47 mg/mL) to achieveisotonicity. Some of the samples included oxidant (methionine 1 mg/mL,EDTA 1 mg/mL, or combination of EDTA 1 mg/mL and methionine 1 mg/mL).Each formulation (see Table below) was filled in a 10R glass vial sealedwith a rubber stopper and an aluminium cap.The solutions were incubated at 40° C. at 75% RH, with samples taken atday 30 (1 month), and day 90 (3 months).Samples were evaluated by HPLC to determine the amount of intact peptideremaining at the various time points, by methods well known in the art,comparing the % Area of the intact peptide peak on the sampling day vs.% Area on Day 0.The HPLC method used an Agilent 1100 instrument. The mobile phases whereHPLC Buffers A (A=0.1% TFA in water) and B (B=0.1% TFA in acetonitrile)with the gradient 20 to 30% B in 40 min, then 30 to 60% B in 15 min,then 60 to 20% B in 1 min and then 20% B for 10 min at flow rate 0.5mL/min. The Zorbax 300SB C18, 3.0×150 mm, 3.5 μm, 300 Å column was attemperature 25 with UV detection at 214 nm. The injection volume was 15μLThe results are shown in the following Table.

TABLE 6 Initial Peptide Peptide peptide conc. conc. Sample Isotonicityconc. (mg/mL) (mg/mL) Number Buffer pH agent Antioxidant (mg/mL) at 30days at 90 days 1 Citrate 6 NaCl No 0.186 0.187 0.182 2 Citrate 5.65NaCl No 0.187 0.187 0.182 3 Citrate 5.8 NaCl No 0.187 0.187 0.182 4Citrate 5 NaCl Methionine 0.187 0.183 0.162 5 Citrate 5.5 NaClMethionine 0.187 0.187 0.171 6 Citrate 6 NaCl Methionine 0.186 0.1870.181 7 Citrate 6 NaCl EDTA 0.187 0.188 0.183 8 Citrate 6 NaClMethionine 0.187 0.187 0.182 and EDTA 8 placebo Citrate 6 NaClMethionine 0.000 0.000 0.000 and EDTA 9 Citrate 5 Mannitol No 0.1870.186 0.173 10 Succinate 6 Mannitol No 0.188 0.186 0.180 11 Succinate 5NaCl No 0.186 0.187 0.183 12 Succinate 5.2 NaCl No 0.187 0.188 0.184 13Succinate 5.65 NaCl No 0.187 0.188 0.183 14 Succinate 6 NaCl No 0.1870.187 0.182 15 Succinate 5 NaCl Methionine 0.187 0.186 0.182 16Succinate 5.2 NaCl Methionine 0.187 0.185 0.182 17 Succinate 5.65 NaClMethionine 0.187 0.186 0.180 18 Succinate 6 NaCl Methionine 0.187 0.1880.181 19 Succinate 5 Mannitol No 0.188 0.186 0.176 20 Succinate 6Mannitol No 0.187 0.099 0.124 21 Succinate 5 Mannitol Methionine 0.1870.180 0.012 21 placebo Succinate 5 Mannitol Methionine 0.000 0.000 0.00022 Succinate 6 Mannitol Methionine 0.188 0.023 0.174 23 Acetate 5.2 NaClNo 0.186 0.186 0.183 24 Acetate 5.65 NaCl No 0.187 0.187 0.184 24placebo Acetate 5.65 NaCl No 0.000 0.000 0.000

Conclusion

FE 202767 showed good stability in citrate and acetate buffers in the pHrange tested after 30 days in each condition. It was also very stable insuccinate buffer at pH 5.0 to 5.65; however, there was significantdegradation after 30 days in some pH 6.0 succinate samples (sample 20,22). The presence or absence of antioxidant seemed unimportant on a 30day timescale.FE 202767 also showed good stability in citrate and acetate buffers inthe pH range tested after 90 days in each condition, with the bestresults being shown at the upper end of the pH range (e.g. between pH5.5 and 6, see samples 1 to 6). It was also stable in succinate bufferat pH 5.0 to 5.65 after 90 days. The 30 and 90 day results for samples21 and 22 suggest a mix up in analysis.Again, the presence or absence of antioxidant seemed unimportant on a 90day timescale.The results indicate that NaCl is a better isotonicity agent thanmannitol.As indicated above, a formulation suitable for nasal administration isexpected to be of pH between 5.0 and 6.0, include the minimum number ofreagents (e.g. no anti-oxidant). It is also preferred that theformulation is room temperature stable. Example 8 demonstrates thatformulations along the lines above may be suitable for nasaladministration, because they have appropriate pH and are roomtemperature stable without requirement for anti-oxidant or otheradditives that might adversely affect the nasal mucosa.

Example 9—Formulation of FE 202767 with Citrate/Phosphate Buffer

FE 202767 (Ferring) was synthesised by the method set out inWO2009/122285. The formulation set out in the following table was madeup by similar methods to those set out in Examples 4 and 5 above.

TABLE 7 Component Amount per mL Function carba-1-[4-FBzlGly7]dOT 0.7 mgActive ingredient (FE 202767) Sodium phosphate dibasic 3.24 mg Bufferingagent dihdrate Citric acid monohydrate 1.43 mg Buffering agent NaCl 7.5mg Isotonicity agent HCl q.s. adjust to pH 5.5 pH adjustment NaOH q.s.adjust to pH 5.5 pH adjustment Water for Injection Adjust to 1 mLSolventThe composition is suitable for nasal administration.

Optionally, an antioxidant (e.g. methionine at a concentration of 1.0mg/mL may be included in the formulation).

1-37. (canceled)
 38. A stable aqueous pharmaceutical compositioncomprising a pharmaceutically active compound of formula (II):

or a pharmaceutically acceptable salt thereof; wherein the compositioncomprises: 2 mg/mL of the compound of formula (II) or a pharmaceuticallyacceptable salt thereof; a citrate-phosphate buffer; and an isotonicityagent; wherein the pH of the composition is from 5.4 to 5.65; andwherein, after storage at 40° C. for 176 days, greater than 95% of thecompound of formula (II) or pharmaceutically acceptable salt thereofremains in the composition.
 39. The composition of claim 38, wherein theisotonicity agent is NaCl.
 40. The composition of claim 38, wherein thepH of the composition is 5.5.
 41. A kit, comprising: the stable aqueouspharmaceutical composition of claim 38; and a container for thecomposition.
 42. A method of treating one or more conditions selectedfrom compromised lactation conditions, labor induction impairment,uterine atony conditions, excessive bleeding, inflammation, pain,abdominal pain, back pain, male and female sexual dysfunction, irritablebowel syndrome (IBS), constipation, gastrointestinal obstruction,autism, stress, anxiety, depression, anxiety disorder, surgical bloodloss, post-partum haemorrhage, wound healing, infection, mastitis,placenta delivery impairment, osteoporosis, comprising administering acomposition according to claim 38 to a patient in need thereof.
 43. Amethod for diagnosis of one or more conditions selected from cancer andplacental insufficiency, comprising administering a compositionaccording to claim 38 to a patient in need thereof.
 44. of treating oneor more conditions selected from uterine atony and excessive bleedingfollowing vaginal delivery comprising administering a compositionaccording to claim 38 to a patient in need thereof.